XB-IMG-74089
Xenbase Image ID: 74089
|
Fig. 5. Tbx2 downregulates the expression of Gremlin and Hey1 to control pronephric morphogenesis. (A) Diagram of the in vitro kidney induction assay. RA, retinoic acid. (B-N) Four-cell stage Xenopus embryos were injected in the animal pole region with Tbx2 mRNA (100 pg) or Tbx2 MO (10 ng) and then the animal explants were dissected at stage 9.5 and processed for in vitro kidney induction. Subsequently, the animal cap tissues were observed for morphology (B-E), sectioned for in situ hybridization with a Pax2 antisense probe (F-I) and immunohistochemistry with the tubule-specific antibody 3G8 (J-M) or subjected to RT-PCR analysis (N). W, whole embryo; AC, uninjected animal caps without RA and activin treatment; (–), uninjected animal caps with RA and activin treatment; –RT, negative control without reverse transcriptase; ODC, ornithine decarboxylase loading control. Arrowheads (G,K,I,M) indicate Pax2 expression or 3G8 staining in the induced animal caps. (O-R) Embryos injected with Tbx2 (200 pg), Tbx2-GR (200 pg), Tbx2 MO (10 ng) or Tbx2δC-GR (200 pg) were subject to in situ hybridization with a Hey1 antisense probe. Arrowheads indicate Hey1 expression in the pronephros. Embryos in P,R were treated with DEX from stage 22 to 32. (S-U) Impaired expression of Pax2 caused by Tbx2 was restored by co-injection of Gremlin or Hey1. Embryos injected with Tbx2 mRNA (200 pg) with or without Hey1 (100 pg) or Gremlin (10 pg) were subject to in situ hybridization for Pax2 at stage 35. n, total number of embryos analyzed; the percentage of embryos showing the phenotype illustrated is indicated. Left and right parts of each panel show the injected and uninjected control sides of embryos, respectively. Image published in: Cho GS et al. (2011) Copyright © 2011. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
Image source: Published Larger Image Printer Friendly View |